Human CCL24/Eotaxin-2/MPIF-2 Quantikine ELISA Kit Summary

Product Summary

The Quantikine Human Eotaxin-2 Immunoassay is a 4.5 hour solid phase ELISA designed to measure Eotaxin-2 levels in cell culture supernates, serum, and plasma. It contains E.coli-expressed recombinant human Eotaxin-2 and antibodies raised against the recombinant factor. Results obtained for naturally occurring human Eotaxin-2 showed linear curves that were parallel to the standard curves obtained using the kit standards. These results indicate that this kit can be used to determine relative mass values for natural human Eotaxin-2.

Preparation and Storage

Background: CCL24/Eotaxin-2/MPIF-2

Eotaxin-2, also named myeloid progenitor inhibitory factor 2 (MPIF-2), is a member of the CC chemokine subfamily and is designated CCL24. Eotaxin-2 is constitutively expressed in the jejunum and spleen. It can also be induced in the lung by allergen challenge and IL-4. Mouse Eotaxin-2 precursor protein shares approximately 58% amino acid sequence identity with human Eotaxin-2.

Assay Procedure

Refer to the product datasheet for the complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.

1.     Prepare all reagents, standard dilutions, and samples as directed in the product insert.

2.     Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

3.     Add 50 μL of Assay Diluent to each well.

4.     Add 50 μL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.

5.     Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.

6.     Add 100 μL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours.

7.     Aspirate and wash 5 times.

8.     Add 100 μL Substrate Solution to each well.

9.     Add 100 μL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.